Development of human iPSC-derived hepatocytes for drug discovery, translational research and toxicity testing

Hepatocytes comprise over 80% of the liver mass and are responsible for most of its functions, including lipid and glucose metabolism, storage of macronutrients, secretion of plasma proteins, detoxification and xenobiotic metabolism. Due to this vast spectrum of functionality, in vitro hepatocytes have been used in both toxicology research and to study debilitating genetic diseases and life-threatening conditions, such as alpha-1-antitrypsin deficiency and MAFLD. Despite the growing burden of liver diseases, treatments remain limited due to challenges with traditional models; e.g. primary human hepatocytes are scarce and difficult to maintain, immortalised cell lines lack key liver functions and animal models present interspecies differences.
Here, we used opti-ox™, a deterministic cell programming technology, to generate consistent, scalable human induced pluripotent stem cell (hiPSC)-derived hepatocytes and demonstrated their suitability for both research and disease modelling (ioHepatocytes Discovery Research), and toxicology (ioHepatocytes Toxicology).

ioHepatocytes display expected cobblestone morphology with distinctive nuclei and well-defined borders. Cells express key pan-hepatocyte markers including ALB, HNF4A, ASGR1 and SERPINA1, and present a transcriptomic signature similar to primary human hepatocytes. Additionally, ioHepatocytes show a high degree of consistency when derived from iPSCs with opti-ox.

ioHepatocytes Discovery Research secrete albumin, store glycogen and accumulate lipids as well as forming bile canaliculi in vitro, demonstrating suitability for modelling metabolic syndromes. ioHepatocytes Toxicology show expression of phase I, II and III drug metabolism genes and have functional cytochrome P450 enzymes (CYP3A, CYP2B6 and CYP1A2). Finally, when challenged with compounds known to cause drug induced liver injury, ioHepatocytes Toxicology shows a similar toxic response to primary human hepatocytes, indicating prospective use for ioHepatocytes Toxicology in the prediction of hepatotoxicity.
ioHepatocytes offer a highly consistent, functional human hepatocyte model, with enhanced scalability to mitigate some of the pitfalls and challenges faced with existing technologies.